. 21
Innovative Research in Engineering Sciences Vol 3(1), 21-25 (2017)
Journal of Innovative Research in Engineering Sciences
ISSN(2476-7611)Journal homepage:
www.Joires.com
Evaluation of The commercial Bacillus thuringiensis (BT) products
Sasha Vaziri1*, Dr. Mahmoud Shojaei2 1
Islamic Azad University, Science and Research Branch of Tehran Agriculture College and Natural Resources Group,Tehran,Iran.
2Agricultural Engineering - Agricultural Entomology, Tehran, Iran.
Abstract: Bacillus thuringiensis (BT) is the most widely used biological insecticide. This bacterium makes protein crystals during sporulate. In this study, effectiveness commercial formulations of Bacillus thuringiensis were tested against 3 moth larvae insect. Concentrations 0.125, 0.062, 0.031, 0.015, 0.25, 0.5, 1, 2 gram and 1.25, 0.625 and 0.315 gram were provided from Bacillus thuringiensis and were effected on Triboliom. The results showed that the ratio of 250 to 500 gram in 100 liters for every Petri of treatment and the amount of 5 gram flour is caused death of the insect.
Keywords: Bacillus thuringiensis, commercial evaluation, Bt products
1. Introduction
Chemical combat has a long history against insects. Farmers started use of the chemicals for pests control in mid-year (1800). It is not surprising that developing the use of insecticides occurred by the Advancement of chemistry Science. There are many disadvantages by using of chemical insecticides. First step, at using of chemical compounds separately cause resistance of
species children toward elective evaluation from pests on children that cause their resistance .For
example, the strains (housefly sepsis Musca domesticae) for contrast with that developed a specific insecticide around the world. The second problem some was about some of pesticides the effect of recombinant species with devastating results were caused, remain devastating effects. Removing unwanted vermin cause will increase other secondary pests. The third worry is related to poisoning arising some of pesticides that result to releasing chemical poisons at environment [1].
In total, presenting of these disadvantages can provide a strong reason to find new ways to control the pests. Like all creatures, insects are susceptible to pathogenic pathogens, such as microorganisms (bacteria, Fungi, Protozoa and viruses)[2].
Bacillus thuringiensis is the most widely used biological insecticide and is a positive-warm aerobic bacterium that was found at the nature. This bacteria makes toxic protein crystals during sporulate and different toxin produced by different strains of Bacillus. The most important toxin was named Dlta_Andotoxin which are used as the main material all of formulations, nowadays. This, in turn, toxin often have formed from proteins were named, Cry visit tocsin. The activity insecticide of Bacillus thuringiensis is at the effect of Cry visit tocsin that act very selective
for insect. Poison crystal is to shape of 2 pyramidal, but it can be on a cubic shape and even be rectangular. There is three factors, for the effect of Bacillus thuringiensis on pests insect and These factors include the proper PH, protease for digestion and break down protein crystals [3].
Commercial formulation include Spore and crystal that should they were eaten by insects. Digestive systems of insect become paralyzed, after eating B.T., but its death was occurred after some days. Bacteria don't increase outside of the body of host and they aren't permanent on the level of plant. crystal is very sensitive to sunlight and broken. Several strains Bacillus thuringiensis were isolated are effective on lepidopteron, flies and beetles. at recent years active strains isolated on against Hymenoptera and lepidopteron [4].
Koorstaci strain has the highest consumption and on the lepidopteron in different agriculture is effective. In Iran, 90 percent of WP powder (Bachtospeindy Paul)has recommended against Witch Phyllophagous and gypsy moth [5].
Bacillus thuringiensis gene as a controller of insects in plants is an interesting phenomenon. Deltol endo-toxin gene is transferred to corn, tomatoes, potatoes, cotton, tobacco and canola. These plants are resistant against pests susceptible to Bacillus thuringiensis. In Iran Bacillus thuringiensis have been investigated with different brand names on various pests [6-8].
2. Materials and Methods
In this article, we investigate the effect of Bacillus thuringiensis Bacteria on moth wheat flour insect (larvae 3) in vitro, and for this purpose should foster moth wheat flour insect in disposable plastic cups and should provide 10 glasses and we pour in every glass one third wheat flour and then we place inside of that many wheat flour moth adult and put net on glass next
Article Info
Received: 17 December 2016 Accepted: 02 March 2017
we close with a cache. Next, on a label, note your first name, surname and required temperature, at the end you put glasses inside incubator at 26 degree. After, examine them every week and you be informed of conditions and position of foster. in continue, you should provide 200 gram of commercial productions of Bacillus thuringiensis Bacteria [9-14].
3. Methodology
The aim of this study is to evaluate Bacillus thuringiensis toxin on third instar larvae 3 of flour moth (Ephestiakuehniella) in vitro. In this study, society and statistical samples consist of flour moth stages of egg, larva, pupa, adult and Bt poison and independent variable consist of counting time of mortality due to different concentrations of Bacillus thuringiensis and dependent variables include of tested insect, prepared concentrations and mortality rate[15-19].
In order to foster of Butterfly flour in Laboratory conditions, at the first prepared 10 Petri and place 5 gram of wheat flour in each petri, flour of each petri was measured by a digital scale and then were placed inside petries. In tests concentration of BT is consider 0.125, 0.062, 0.031, 0.015, 0.25, 0.5, 1, 2 gram net and 1.25, 0.625, 0.315 gram diluted. Thus[20,21], on the digital scale sheet was placed a piece of silver paper and some BT were poured on silver paper and this way was repeated 5 times. Because, from 10 Petri, 5 of them were for control samples (without pesticides) and 5 Petri was related to repeat, means the sample containing toxin. Some BT was poured on flour inside of repeated Petri and flour and toxin blended completely by SWAP . After blending flour of 5 Petri
to toxin should isolate third age of larvae from adult insects and flour. Samples go out from incubator and were on the table. In order to preventing from escape of full insects, the samples were placed inside of freezer to adult insects be numb. after 2 or 3 minutes we remove glasses from freezer and put them on the table. Separation of larvae was done. At the first, adult insects were removed from inside of glass by SWAP and next were placed inside of Petri. Separation of larvae was done. first way was done manually and by using of 2 SWAP that their tip were a little wet, larvae 3 isolated from flour and other larvae. Then, sieve including flour and larvae were placed under pressure of water. By helping water pressure larvae isolated from flour and using forceps were removed larvae and were placed in petri dishes. Thus, larvae 3 were isolated and were placed on the flour. on every control Petri were placed 5 gram flour without poison BT and 10 larvae 3 and on every repeat petri 5 gram flour mixed to toxin of BT, were placed 10 larvae 3.then the door of petri attached with adhesive tape, then on the Petri door a number, including the poison name, insect scientific name and temperature of testing. Petri dishes were placed in an incubator at a temperature of 27 degrees. After doing test, related results to mortality and the impact of Bt were observed and recorded in different time intervals[22-24].
4. Analyzing results
In order to answer the question, whether the Bacillus thuringiensis bacteria cause death of insects or insect survives, as well as we examine if the bacteria cause mortality, how much is the mortality rate? The results of eleven tests are reported in Table 1.
Table 1. The Results of analyze test of variance related to different concentrations
Meaningful Test F
Average of squares Total of squares
Degree of freedom Change
resource concentration
test
0.04∗∗
5.701 82.283
740.555 9
treatment 0.125
1
14.421 577.235
40 error
1317.79 49
total
0.000∗
10.930 125.672
1131.064 9
treatment 0.062
2
11.497 444.648
40 error
1575.712 49
total
0.000∗
14.816 87.125
784.215 9
treatment 0.031
3
5.881 235.34
40 error
1019.455 49
total
0.001∗
11.405 126.839
1231.553 9
treatment 0.015
4
11.998 479.942
40 error
1711.495 49
total
0.009∗
8.74 117.169
1054.528 9
treatment 0.25
5
13.406 536.25
40 error
159.788 49
total
0.000∗
9.724 106.058
954.528 9
treatment 0.5
6
10.906 436.25
40 error
1390.778 49
total
0.009∗
8.994 109.361
984.257 9
treatment 1
7
12.158 486.345
40 error
1470.602 49
total
0.000∗
12.933 123.501
1201.512 9
treatment 2
8
10.322 412.895
40 error
1614.407 49
. 23 1691.818 49 total 0.000∗ 8.64 115.836 1042.526 9 treatment 0.625 10 13.406 536.25 40 error 1578.776 49 total 0.000∗ 9.653 125.058 1125.526 9 treatment 0.315 11 12.956 518.25 40 error 1643.776 49 -
As you see at the above table Null hypothesis based on Bacillus thuringiensis bacteria with the consumption range of BT 0.125, 0.062, 0.031, 0.015, 0.25, 0.5, 1, 2 gram net and 1.25, 0.625 and 0.315 gram diluted, don't cause the death of insect and for following concentration at the meaningful level was rejected : 0.125 (F=5.701, P<0.05), 0.062 (F=10.930, P<0.01(, 0.031 (F=14.816, P<0.01), 0.015 ( ( F=11.405, P<0.01), 0.25 (F=8.74, P<0.01), 0. (F=9.724, P<0.01), 1 (F=8.994, P<0.01), 2 (F=12.933, P<0.01), 1.25
(F=11.147, P<0.01), 0.625 (F=8.64 , P<0.01), 0.315 el (F=9.653 , P<0.01). on the other hand, Bacillus thuringiensis bacteria with the range of consumption BT : 0.125, 0.062, 0.031, 0.015, 0.25, 0.5, 1, 2 gram net and 1.25, 0.625 and 0.315 gram diluted for every repetition and 5 gram flour cause the death of insect.
At the following table due to the meaningful of different tests, we have specified difference between Examined characteristics at different repetition .
Table 2. Difference between the averages of Examined characteristics at different repetition
eleventh treatment tenth treatment ninth treatment eighth treatment seventh treatment sixth treatment fifth treatment forth treatment third treatment second treatment First treatment 0.6589a 0.5849a 0.5126a 0.7215ab 0.5274a 0.7245a 0.5232a 0.6162a 0.9152ab 0.6157a 0.7233a 1 repeat 0.815a 0.5658a 0.5746a 0.5746a 0.5862a 0.6254a 0.6254a 0.7354a 0.8457ab 0.6632a 0.7033a 2 repeat 0.748a 0.7225ab 0.7685ab 0.7584ab 0.6845ab 0.7542ab 0.6562a 0.7486a 0.7146a 0.7856a 0.65a 3 repeat 0.8456ab 0.8131ab 0.8152ab 0.8259ab 0.7154ab 0.8489ab 0.7725ab 0.8312ab 0.8312ab 0.8338ab 0.7433b 4 repeat 0.6584a 0.8183ab 0.8254ab 0.8875ab 0.8174ab 0.6742bc 0.6456bc 0.7567bc 0.9567ab 0.9567ab 0.8367b 5 repeat
Those written letters at the top of numbers means that the numbers have common letters are significantly different rate and the numbers that are not identical letters are significantly different.
The Results average of mortality rate of controls and treatments at various concentrations and different times, respectively, in Tables 3 and 4 have been reported.
Table 3. The results of average of percentage mortality of controls at different times After (hours) test 264 240 216 192 168 96 72 48 24 16% 14% 10% 8% 6% 0 0 0 0 First 16% 14% 10% 8% 6% 0 0 0 0 second 16% 14% 10% 8% 6% 0 0 0 0 third 18% 14% 8% 6% 4% 0 0 0 0 forth 16% 12% 8% 6% 4% 0 0 0 0 fifth 18% 14% 8% 6% 4% 0 0 0 0 sixth 18% 14% 8% 6% 2% 0 0 0 0 seventh 18% 14% 8% 6% 4% 0 0 0 0 eighth 16% 12% 8% 6% 4% 0 0 0 0 ninth 18% 14% 10% 8% 6% 0 0 0 0 tenth 18% 14% 8% 6% 4% 0 0 0 0 eleventh
Conclusion
For reviewing this matter that whether Bacillus thuringiensis bacteria cause mortality and also we review, if bacteria cause mortality, how much is percentage of mortality? We use from analysis of variance that all tests are meaningful and was used from test of comparison of the average LSD in order to used average comparisons. Results of 11 tests are as follows.
Bacillus thuringiensis Bt by the rang of consumed BT 125/0 gram net for each iteration and 5 grams of flour caused the death of the insect. In other words, test of hypotheses at significant level of 0.04 (F = 5.701, P <0.05) refuted. Also, test of average comparison LSD has many differences between repeat 5 with repeat 3. Bacillus thuringiensis bacteria by the range of consumed BT 0.062 gram net for every repeat and 5 gram flour cause the death of insect. In other words, test hypotheses significant level of 0.000 (F = 10.930, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat 5 with repeat 1. Bacillus thuringiensis bacteria by the range of consumed BT 0.031 gram net for every repeat and 5 gram flour, cause the death of insect. In other words, test hypotheses significant level of 0.000 (F = 14.816, P <0.01) refuted. In other words, test hypotheses significant level of 0.001 (F = 11.405, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat 4 with repeat 1. Bacillus thuringiensis bacteria by the range of consumed BT 0.25 gram net for every repeat and 5 gram flour cause the death of insect. In other words, test hypotheses significant level of 0.009 (F = 8.740, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat 1 with repeat 4. Bacillus thuringiensis bacteria by the range of consumed BT 0.5 gram net for every repeat and 5 gram flour, cause the death of insect. In other words, test hypotheses significant level of 0.000 (F = 9.724, P <0.01) reject the result. Also, test of average comparison LSD has many differences between repeat 4 with repeat 2. Bacillus thuringiensis bacteria by the range of consumed BT 1 gram net for every repeat and 5 gram flour, cause the death of insect. In other words, test hypotheses significant level of 0.009 (F = 8.994, P <0.01) reject the result. Also, test of average comparison LSD has many differences between repeat 1 with repeat 5. Bacillus thuringiensis bacteria by the range of consumed BT 2 gram net for every repeat and 5 gram flour, cause the death of insect. IN other words, test hypotheses significant level of 0.000 (F = 12.933, P <0.01) reject the result. Also, test of average comparison LSD has many differences between repeat 5 with repeat 4. Bacillus thuringiensis bacteria by the range of consumed BT 0.062 gram net for every repeat and 1/25 gram flour cause the death of insect. In other words, test hypotheses significant level of 0.007 (F = 11.147, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat
and 0.625 gram flour cause the death of insect. In other words, test hypotheses significant level of 0.000 (F = 8.640, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat 5 with repeat 2. Bacillus thuringiensis bacteria by the range of consumed BT 0.315 gram diluted for every repeat and 5 gram flour, cause the death of insect. In other words, test hypotheses significant level of 0.000 (F = 9.652, P <0.01) refuted. Also, test of average comparison LSD has many differences between repeat 4 with repeat 1 and 5.
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